去甲万古霉素生产菌株Amycolatopsis orientalis遗传操作体系的建立

The Establishment of Genetic Manipulation System for Norvancomycin Producing Strain Amycolatopsis orientalis

为了对去甲万古霉素高产菌中的功能基因进行研究，以整合型pZMW为载体，考察了去甲万古霉素生产菌东方拟无枝酸菌属间结合转移限制因素，优化3个关键限制因素：产孢培养基为YD，起始孢子量控制在1×10^8～4×10^8／反应，抗生素覆盖时间为16～17h为宜。建立了东方拟无枝酸菌生产菌株NCPC2-48的操作方法和培养条件，使体内分析该菌的生物合成及调控基因的功能成为可能，并为建立其他放线菌遗传操作体系提供了参考。

In order to study the functional genes in Amycolatopsis orientalis, the key constraints factors of conjugation from E. coli to A. orientalis were studied by the integrative plasmid pZMW. Three key restricted factors were optimized： YD spore-producing medium, preliminary spore amount was controlled at 1 × 10^8- 4 × 10^8/reaction, incubation time before covered by antibiotics at suitable at 16 - 17 hours. A genetic manipulation system for the industrial strain A. orientalis NCPC 2-48 was developed which would make the analysis of the functions of biosynthetic and regulation genes of the strain in vivo possible, and provided references for the establishment of the genetic manipulation system of the other Actinomycetes.